Triple quadrupole lc ms ms 6410 system

Metabolites were extracted from rice cells (40 mg) or rice leaves (80–100 mg) and measured on a triple quadrupole LC-MS/MS 6410 system (Agilent Technologies, Santa Clara, CA, USA) equipped with a Zorbax SB-C18 column (50 × 2.1 mm ID, 1.8 μm, Agilent Technologies) as described previously23 (link). Amount of each metabolite was estimated using synthetic standards (CoP, FP) or purified chemicals momilactone A (MoA) and momilactone B (MoB).

The OS isolates were centrifuged and diluted 100-fold before applying 10 μL on a triple quadrupole LC-MS/MS 6410 system (Agilent Technologies, Santa Clara, CA, USA) equipped with a Zorbax SB-C18 column [50 × 2.1 mm ID, 1.8 μm, Agilent Technologies]. MS was set to operate in negative electrospray ionization (ESI) mode and mass scan data were collected in m/z 150–1000 range. Sample separation was achieved with solvent A [0.1% (v/v) formic acid in water] and solvent B [0.1% (v/v) formic acid in acetonitrile] in time (min)/B (%) gradient program: 0/5, 0.5/5, 2/40, 6/40, 10/95, 15/95, 16/5, 20/5. Constant flow rate of 0.4 mL/min was applied to chromatographic column placed in thermostatic chamber controlled at 40 °C. Additional MS variable parameters were used as follows: fragmentor 135V, dwell 200, and 7V cell accelerator voltage. Synthetic FAC (N-linolenoyl-L-Gln) was used to estimate FAC content in individual OS isolates, which was then expressed as N-linolenoyl-L-Gln equivalents.